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. Author manuscript; available in PMC: 2017 Feb 23.
Published in final edited form as: Cell Rep. 2017 Feb 7;18(6):1444–1457. doi: 10.1016/j.celrep.2017.01.023

Figure 6. TELO2 controls mTORC1:substrate association and explains MNK regulation of mTORC1.

Figure 6

(A–C) Dox-inducible, HA-Raptor expressing HEK293 cells were transfected with empty vector (PC3), Flag-DEPTOR (A), Flag-TELO2 (B), or Flag-DDB1 (C), lysed, and subjected to Flag-IP followed by immunoblot analysis.

(D) Flag-mTOR IP of HEK293 cells transfected with empty vector (PC3), untagged TELO2, or untagged TELO2 and DEPTOR combined. Co-IP of relevant proteins was assessed by immunoblot.

(E) Dox inducible, HA-Raptor expressing HEK293 cells were transfected with ctrl (-), TELO2, or MNK1-targeting siRNA. HA-Raptor IP was performed and associated proteins were analyzed by immunoblot.

(F) HEK293 cells were transfected with ctrl (-) or MNK1 and 2-targeting siRNA, transfected with empty vector or flag-TELO2. Cell lysates were assessed by Flag-TELO2 IP and immunoblot.

(G) HeLa cells were transfected with ctrl (-) or MNK1-targeting siRNA followed by transfection with empty vector (PC3), Flag-TELO2, or Flag-MNK1 (T344D). Lysates were analyzed by immunoblot for markers of mTORC1/MNK activation.

Quantitation represents the average of two (A, C, D) or three (B, E, F, G) independent experiments normalized by setting empty vector (PC3) control to 1 (A–C), setting TELO2 + PC3 control to 1 (D), or by setting ctrl siRNA samples to 1 (E–G).